Metal chelators and methods of their use

ABSTRACT

Metal chelators of Formula I and Formula II are disclosed: 
     
       
         
         
             
             
         
       
         
         
           
             or a pharmaceutically acceptable salt thereof. 
           
         
       
    
     Also disclosed are metal chelator-targeting moiety complexes, metal chelator-targeting moiety-metal conjugates, kits, and methods of their preparation and use in diagnosis and/or treatment of diseases and conditions, including, inter alia, cancer and thrombosis.

CROSS REFERENCE TO RELATED APPLICATIONS

The application is a divisional of U.S. application Ser. No. 11/659,601filed Feb. 6, 2008 which claims priority to U.S. Application No.60/600,253 filed Aug. 10, 2004 and to U.S. Application No. 60/603,781filed Aug. 23, 2004, the entire disclosures of which are incorporatedherein by reference.

STATEMENT OF FEDERALLY SPONSORED RESEARCH

The U.S. Government may have certain rights in the invention describedherein, which was made in part with funds from NIH Contract No.263-02-D-0053.

FIELD OF THE INVENTION

The present invention relates to metal chelators, metalchelators-targeting moiety complexes, metal chelators-targetingmoiety-metal conjugates, kits, and methods of their preparation and usein diagnosis and/or treatment of diseases and conditions, including,inter alia, cancer and thrombosis.

BACKGROUND OF THE INVENTION

Direct imaging of many biochemical processes is now practicable throughthe use of radiopharmaceuticals targeted towards specificdisease-associated molecular targets. This was made possible by thediscoveries in the field of disease related changes in cellularcommunication and metabolism, especially in cancer. To support these newdiagnostic applications, methods for linking radioisotopes to theappropriate targeting biomolecules were required to replace the simplemetal chelates and ions used previously. In the 1980s and 1990s, methodswere developed for labeling biomolecules, especially monoclonalantibodies, with radionuclides such as technetium-99m (^(99m)Tc) andindium-111 (¹¹¹In). In its most developed form, this typically entailedcovalent attachment of a bifunctional chelator to a protein, followed bylabeling with the radiometal, or even synthesis of a radiometalbifunctional chelate that was subsequently attached to the protein. Asrecognition grew in the 1980s that monoclonal antibodies were too largeto offer ideal pharmacokinetics, focus shifted onto smaller molecules,such as antibody fragments and especially smaller peptides, targetedtowards receptors present in lesions such as tumors and thrombi.Examples include radiolabeled octatreotide (selective for somatostatinreceptors expressed by tumors of the endocrine system such asparagangliomas and neuroblastomas), bombesin (receptors for this peptideare expressed by small-cell lung carcinomas), and α-melanocytestimulating hormone (expressed by melanomas).

Although the transition to smaller molecules brought with it theopportunity to use peptides produced by solid phase peptide synthesis(SPPS) rather than proteins of biological origin, the same methods wereused to label them as had been used to label antibodies. These methodshave several disadvantages, which are more problematic with smallpeptides than with large proteins. The most suitable sites forattachment of a bifunctional chelator in most peptides are the ε-aminogroups of lysine residues and the N-terminus, because they are veryreactive nucleophiles and form very unreactive covalent links with thechelator. If there is more than one lysine in the peptide chain, thesite of modification becomes uncertain. For instance, if the peptide hastwo lysines, together with the N-terminus these will present threepossible sites for conjugation, hence forming as many as eight productswhen treated with an active-ester-containing bifunctional chelator orradiolabeled bifunctional chelate. Each of these products will have adifferent biodistribution and different affinities for the target (someof which may have lost all target affinity) and such a mixture is notacceptable for clinical use. Moreover, one or more of the lysines may beessential to the biological activity of the peptide. A simple solutionhas been to incorporate the chelator, or a radiolabeled chelate ororganic prosthetic group, as the last step of SPPS. This, however, hasthe limitation that the chelator has to be at one end of the peptidechain, which is frequently essential to the biological activity of thepeptide.

The state of the art in linking radiometals to peptides encompasses anumber of approaches. Some have the advantage of incorporating the metalbinding sequence during SPPS, and others have the advantage ofincorporating chelators that are specifically designed for theparticular metal. Few, however, have both of these advantages. Forexample, technetium-chelating amino acid sequences such as gly-gly-cysare incorporated during SPPS or recombinant protein production, but thissequence is not ideal for its purpose, and merely represents the bestthat can be achieved for chelating the TcO³⁺ core using “standard” aminoacids (i.e. those coded through tRNAs). Likewise, polyhistidinesequences, such as hexahistidine, can be incorporated during SPPS, butagain they merely represent the best sequence of coded amino acidsachievable for chelating the Tc(CO)₃ ⁺ core. Conversely, the synthetictechnetium ligand hynic (hydrazinonicotinamide) probably represents themost convenient and efficient labeling system to date for use with^(99m)Tc, but it has so far only been used by conjugating it to apre-formed peptide, with all the associated problems outlined above. Analternative that offers convenience of labeling is the “direct labeling”method in which antibodies and peptides containing disulfide bonds canbe reduced and labeled with ^(99m)Tc or ¹⁸⁸Re. However, the chemistry ofthese methods is poorly understood, and there are major stability andbiological activity problems as demonstrated by the work of severalgroups world wide with antibodies and somatostatin analogues.

WO 2004/022106 discloses technology that incorporates the metal bindingmoiety during SPPS and incorporates chelators that are specificallydesigned for the particular metal. More specifically, WO 2004/022106discloses metal-chelating precursors, designed to bind specific metallicradionuclides and incorporating a pendant protected (e.g. Fmoc) aminoacid functionality. The chelator is attached to an amino acid beforerather than after SPPS assembly of the peptide chain. Achelator-derivatized amino acid comprises: 1) an optionally protectedprimary or secondary amino group; 2) a carboxylic acid group; 3) achelator group capable of binding a metallic radionuclide. Suitablechelating or metal binding groups may be chosen from several structuresincluding but not limited to the hydrazinonicotinamide group, di- orpoly-thiol groups, macrocyclic ligands incorporating amine, thioether,or phosphine donor groups, or polyaminocarboxylate groups.

As discussed above, there is a continuing need for more versatile andcontrolled approaches to the synthesis of metal conjugates useful, interalia, in the diagnosis and/or treatment of diseases and conditions. Thepresent invention focuses on novel compounds directed to these and otherimportant uses.

SUMMARY OF THE INVENTION

The present invention is directed to metal chelators, metalchelator-targeting moiety complexes, metal chelator-targetingmoiety-metal conjugates, kits, and methods of their preparation and usein diagnosis and/or treatment of diseases and conditions, including,inter alia, cancer and thrombosis. The metal chelators may be used inconventional synthetic methods, inter alia, to form targeting moieties,such as peptides, proteins, and amine-surface functional dendrimers,capable of conjugating diagnostic and/or therapeutic metals.

In one embodiment, the invention is directed to compounds of Formula Iand II:

or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention is directed to complexes,comprising:

-   a. at least one targeting moiety selected from the group consisting    of peptide, protein, and an amine-surface functional dendrimer; and-   b. a residue of a compound of Formula I or Formula II or a    pharmaceutically acceptable salt thereof;

wherein said compound is covalently linked to said targeting moiety.

In other embodiments, the invention is directed to conjugates,comprising:

-   a. at least one targeting moiety selected from the group consisting    of peptide, protein, and amine-surface functional dendrimer;-   b. a residue of a compound of Formula I or Formula II or a    pharmaceutically acceptable salt thereof;

wherein said compound is covalently linked to said targeting moiety; and

-   c. a diagnostic or therapeutic metal.

In other embodiments, the invention is directed to compositions,comprising:

a. a complex comprising:

-   -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   wherein said compound is covalently linked to said targeting        moiety; and        b. a pharmaceutically-acceptable carrier.

In another embodiment, the invention is directed to compositions,comprising:

a. a conjugate, comprising:

-   -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer;    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;        -   wherein said compound is covalently linked to said targeting            moiety; and    -   iii. a diagnostic or therapeutic metal; and        b. a pharmaceutically-acceptable carrier.

In yet other embodiments, the invention is directed to kits fordetecting, imaging, monitoring, or treating a disease or condition in apatient comprising:

-   a. a complex comprising:    -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   wherein said compound is covalently linked to said targeting        moiety;-   b. a therapeutic or diagnostic metal;-   c. an optional pharmaceutically-acceptable carrier; and-   d. instructions for preparing a composition comprising a therapeutic    or diagnostic agent for detecting, imaging, monitoring, or treating    a disease or condition in a patient.

In yet other embodiments, the invention is directed to kits fordetecting, imaging, monitoring, or treating a disease or condition in apatient comprising:

-   a. a conjugate comprising:    -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   iii. a diagnostic or therapeutic metal;        -   wherein said compound is covalently linked to said targeting            moiety;-   b. an optional pharmaceutically-acceptable carrier; and-   c. instructions for preparing a composition comprising a therapeutic    or diagnostic agent for detecting, imaging, monitoring, or treating    a disease or condition in a patient.

In other embodiments, the invention is directed to processes ofsynthesizing peptides or proteins comprising a chelator, said processescomprising the step of:

incorporating into said peptides or proteins a residue of a compound ofFormula I or Formula II or a pharmaceutically acceptable salt thereof.

In other embodiments, the invention is directed to processes ofsynthesizing amine-surface functional dendrimers comprising a chelator,said processes comprising the step of:

incorporating into said amine-surface functional dendrimer a residue ofa compound of Formula I or Formula II or a pharmaceutically acceptablesalt thereof.

In yet another embodiment, the invention is directed to conjugates ofthe peptides, proteins, and amine-surface functional dendrimers producedby the above-described processes.

In other embodiments, the invention is directed to methods of detecting,imaging or monitoring cancer in a patient, comprising the steps of:

-   a. administering to said patient a conjugate described above; and-   b. acquiring an image of a site of concentration of said conjugate    in the patient by a diagnostic imaging technique.

In other embodiments, the invention is directed to methods of detecting,imaging or monitoring thrombi in a patient, comprising the steps of:

-   a. administering to said patient a conjugate described above; and-   b. acquiring an image of a site of concentration of said conjugate    in the patient by a diagnostic imaging technique.

In yet other embodiments, the invention is directed to methods oftreating cancer, comprising the step of:

administering to a patient in need thereof an effective amount of aconjugate described above.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to metal chelators, metalchelator-targeting moiety complexes, metal chelator-targetingmoiety-metal conjugates, and methods of their preparation and use indiagnosis and/or treatment of diseases and conditions, including, interalia, cancer and thrombosis. The metal chelators may be used inconventional synthetic methods, inter alia, to form targeting moieties,such as peptides, proteins, and amine-surface functional dendrimers,capable of conjugating diagnostic and/or therapeutic metals. Thefollowing definitions are provided for the full understanding of termsand abbreviations used in this specification.

DEFINITIONS

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include the plural reference unless the context clearlyindicates otherwise. Thus, for example, a reference to “a compound” is areference to one or more compounds and equivalents thereof known tothose skilled in the art, and so forth.

As used herein, the term “therapeutic agent” refers to an agent that maybe used to prevent cure, alleviate the onset and/or progression of acondition(s), pathological disorder(s) or disease(s).

As used herein, the term “diagnostic agent” refers to an agent that maybe used to detect, image and/or monitor the presence and/or progressionof a condition(s), pathological disorder(s) or disease(s).

A “chelator” is the moiety or group on a reagent that binds to a metalion through the formation of chemical bonds with one or more donoratoms.

As used herein, the phrase “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms that are, withinthe scope of sound medical judgment, suitable for use in contact withthe tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts prepared from pharmaceutically acceptable non-toxic acids,including inorganic salts, and organic salts. Suitable non-organic saltsinclude inorganic and organic acids such as acetic, benzenesulfonic,benzoic, camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic,glutamic, hydrobromic, hydrochloric, isethionic, lactic, malic, maleic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phosphoric, succinic, sulfuric, tartaric acid, p-toluenesulfonic and thelike. Particularly preferred are hydrochloric, hydrobromic, phosphoric,and sulfuric acids, and most preferably is the hydrochloride salt.

As used herein, term “administering” means either directly administeringa compound, complex, conjugate, or composition of the present invention,or administering a prodrug, derivative or analog that will form anequivalent amount of the active compound or substance within the body.

The term “effective amount,” as used herein, refers to an amounteffective, at dosages, and for periods of time necessary, to achieve thedesired result with respect to the therapeutic or diagnostic treatment.

As used herein, the term “subject” or “patient” refers to an animalincluding the human species that is treatable with the compositions,and/or methods of the present invention. The term “subject” or“subjects” is intended to refer to both the male and female genderunless one gender is specifically indicated. Accordingly, the term“patient” comprises any mammal that may benefit from diagnosis and/ortherapy using the metal chelators of the invention.

Metal Chelator Compounds

In one embodiment, the invention is directed to compounds of Formula Iand II:

or a pharmaceutically acceptable salt thereof.

The invention also contemplates the deprotected compound of Formula II,where the fmoc group is not present and is replaced by a hydrogen atom.Thus, the “compound of Formula II,” as used herein, includes thedeprotected analog of the compound of Formula II:

The metal chelator compound of Formula I may be prepared in accordancewith the general synthetic scheme shown in Scheme 1.

The synthesis of the compound of Formula I is completed by fullyalkylating the amines with bromoacetate t-butyl ester, hydrogenation ofthe nitro group, and reaction of the aniline with glutaric anhydride,simultaneously protecting the aniline nitrogen as an amide and providinga terminal carboxylate suitable for conversion to an activated ester.COESY NMR spectra of the carboxylic acid intermediate shows atriplet-pentet-triplet pattern of the glutaryl-linking moiety.

In certain preferred embodiments, a specific enantiomer(S,S-cyclohexyl-R-p-nitrophenylalenyl) is prepared. Aminocarboxylatechelates with this specific enantiomeric backbone are preferred becauseof their superior metal ion complexation stability.

-   1. Wu, C., Kobayashi, H., Sun, B., Yoo, T. M., Paik, C. H.,    Gansow, O. A., Carrasquillo, J. A., Pastan, I., Brechbiel, M. W.:    Stereochemical Influence on the Stability of Radio-Metal Complexes    In Vivo. Synthesis and Evaluation of the Four Stereoisomers of    2-(p-nitrobenzyl)-trans-CyDTPA. Bioorg. Med. Chem. 1997, 5,    1925-1934.-   2. Brechbiel, M. W., Pippin, C. G., McMurry, T. J., Milenic, D.,    Roselli, M., Colcher, D., Gansow, O. A.: An Effective Chelating    Agent for Labeling of Monoclonal Antibody with ²¹²Bi for a-Particle    Mediated Radioimmunotherapy. J. Chem. Soc., Chem. Commun. 1991,    1169-1170.-   3. Milenic, D. E., Garmestani, K., Chappell, L. L., Dadachova, E.,    Yordanov. A., Ma, D., Schlom, J., Brechbiel, M. W. In Vivo    Comparison of Macrocyclic and Acyclic Ligands for Radiolabeling of    Monoclonal Antibodies with ¹⁷⁷Lu for Radioimmunotherapeutic    Applications. Nucl. Med. Biol. 2002, 29, 431-442.

The metal chelator compound of Formula II may be prepared by mixing acompound of Formula I with a commercially-available sample of an9-fluorenylmethoxycarbonyl (FMOC or fmoc) protected lysine to yield acompound of Formula II. The compound of Formula II may be deprotected byany conventional techniques for removing FMOC protecting groups,including, for example, treatment with base such as 20-50% piperidine indimethylformamide (DMF) for about 20 minutes. In the case of incompletefmoc deprotection, a stronger base, such as1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) with 2% piperidine, may beused.

As will be readily understood, functional groups present may containprotecting groups during the course of synthesis. Protecting groups areknown per se as chemical functional groups that can be selectivelyappended to and removed from functionalities, such as hydroxyl groupsand carboxyl groups. These groups are present in a chemical compound torender such functionality inert to chemical reaction conditions to whichthe compound is exposed. Any of a variety of protecting groups may beemployed with the present invention. Protecting groups that may beemployed in accordance with the present invention may be described inGreene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis2d. Ed., Wiley & Sons, 1991.

It should be understood that the compounds of this invention may bemodified by appending appropriate chemical groups to enhance selectivebiological properties. Such modifications are known in the art andinclude those that increase biological penetration into a givenbiological compartment (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

It should also be understood that the compounds of this invention mayadopt a variety of conformational and ionic forms in solution, inpharmaceutical compositions and in vivo. Although the depictions hereinof specific compounds of this invention are of particular conformationsand ionic forms, other conformations and ionic forms of those compoundsare envisioned and embraced by those depictions.

Metal Chelator-Targeting Moiety Complexes

In another embodiment, the invention is directed to complexes,comprising:

-   a. at least one targeting moiety selected from the group consisting    of peptide, protein, and amine-surface functional dendrimer; and-   b. a residue of a compound of Formula I or Formula II or a    pharmaceutically acceptable salt thereof;

wherein said compound is covalently linked to said targeting moiety.

The compound of Formula I may be incorporated at the end of the chain(s)of the peptide, protein, or amine-surface functional dendrimer. Compoundof Formula II may be incorporated either at the end or within thechain(s) of the peptide, protein, or amine-surface functional dendrimer.

In certain embodiments, the targeting moiety is poly(amidoamine)dendrimer (PAMAM), octatreotide, bombesin, or α-melanocyte stimulatinghormone and the residue of a compound of Formula I or Formula II isincorporated at the end of the peptide/dendrimer or the compound ofFormula II is incorporated within the peptide/dendrimer backbone.

In certain embodiment of the complex, the residue of a compound ofFormula I or Formula II is linked to the targeting moiety via a terminalamino group on the targeting moiety, preferably via an N-terminus of thetargeting moiety, a lysine residue on the targeting moiety, or shellamine of an amine-surface functional dendrimer.

In certain other embodiment of the complex, the residue of a compound ofFormula II may be linked to the targeting moiety at any pre-determinedpoint in the sequence of the targeting moiety, including at theN-terminal, between two units of amino acids, or on a lysine residue.When a compound of Formula II is used, a single complex may contain morethan one unit of a compound of Formula II, for example at the N-terminaland between two units of amino acids.

In other embodiments, the invention is directed to processes ofsynthesizing peptides or proteins comprising a chelator, said processescomprising the step of:

incorporating into said peptides or proteins a residue of a compound ofFormula I or Formula II.

In other embodiments, the invention is directed to processes ofsynthesizing amine-surface functional dendrimers comprising a chelator,said processes comprising the step of:

incorporating into said amine-surface functional dendrimer a residue ofa compound of Formula I or Formula II or a pharmaceutically acceptablesalt thereof.

In yet another embodiment, the invention is directed to complexes ofpeptides, proteins, and amine-surface functional dendrimers produced bythe above-described processes.

Typically, the complex will be synthesized prior to chelation of thetherapeutic or diagnostic metal. However, the present inventioncontemplates the chelation of the therapeutic or diagnostic metal to thecompound of Formula I or Formula II or a pharmaceutically acceptablesalt thereof prior to synthesis of the complex.

Preferably, when the targeting moiety is a peptide or protein, it may besynthesized by solid phase peptide synthesis (SPPS), such as describedin Atherton, E., Sheppard, R. C., Solid phase peptide synthesis: apractical approach. IRL Press at Oxford University Press (1989). Anexample is shown below with a compound of Formula I:

The amine-surface functional dendrimers, preferably PAMAM-type, may beprepared by conventional synthetic techniques, such as those describedin Jean M. J. Frechet (editor), Donald A. Tomalia (editor), Dendrimersand Other Dendritic Polymers (New York: John Wiley & Sons, 2002).

The method of the present invention is advantageous over conventionalpost-SPPS conjugation in at least the following ways:

-   1. The method is versatile and flexible: it incorporates the    chelator site-specifically anywhere in sequence in the case of the    chelator compound of Formula II, not just terminally or at one or    more lysines.-   2. It can leave lysines unmodified.-   3. It is economic since no post-SPPS modification is needed.

Metal Chelator-Targeting Moiety-Metal Conjugates

In other embodiments, the invention is directed to conjugates,comprising:

-   a. at least one targeting moiety selected from the group consisting    of peptide, protein, and amine-surface functional dendrimer;-   b. a residue of a compound of Formula I or Formula II or a    pharmaceutically acceptable salt thereof;

wherein said compound is covalently linked to said targeting moiety; and

-   c. a diagnostic or therapeutic metal.

In other embodiments, the invention is directed to compositions,comprising:

a. a complex comprising:

-   -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   wherein said compound is covalently linked to said targeting        moiety; and        b. a pharmaceutically-acceptable carrier.

Suitable diagnostic and therapeutic metals include paramagnetic metalions, gamma-emitting radioisotopes, positron-emitting radioisotopes, andx-ray absorbers.

Suitable paramagnetic metal ions include Gd(III), Dy(III), Fe(III), andMn(II).

Suitable gamma-emitting radioisotope or positron-emitting radioisotopeinclude ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, ^(99m)Tc, ⁹⁵Tc, ¹¹¹In, ⁹⁰Y, ¹⁴⁹Pr,¹⁵³Sm, ¹⁵⁹Gd, ¹⁶⁶Ho, ¹⁶⁹Yb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, and ²¹³Bi, preferably,^(99m)Tc, ¹¹¹In, and ²¹³Bi.

Suitable x-ray absorbers include Re, Sm, Ho, Lu, Pm, Y, Bi, Pd, Gd, La,Au, Au, Yb, Dy, Cu, Rh, Ag, and Ir.

The conjugates of the invention are formed by contacting, preferablywith simple mixing, the desired metal with the desired complex of theinvention.

Compositions

The compounds and the conjugates of the invention may be used as a neatcomposition or as a composition containing at least one pharmaceuticallyacceptable carrier. Generally, the residue of compound of Formula I orFormula II, or a pharmaceutically acceptable salt thereof, will bepresent at a level of from about 0.1%, by weight, to about 90% byweight, based on the total weight of the composition, based on the totalweight of the composition. Preferably, the residue of compound ofFormula I or Formula II or a pharmaceutically acceptable salt thereofwill be present at a level of at least about 1%, by weight, based on thetotal weight of the composition. More preferably, the residue ofcompound of Formula I or Formula II or a pharmaceutically acceptablesalt thereof will be present at a level of at least about 5%, by weight,based on the total weight of the composition. Even more preferably, theresidue of compound of Formula I or Formula II or a pharmaceuticallyacceptable salt thereof will be present at a level of at least about10%, by weight, based on the total weight of the composition. Yet evenmore preferably, the residue of compound of Formula I or Formula II or apharmaceutically acceptable salt thereof, will be present at a level ofat least about 25%, by weight, based on the total weight of thecomposition.

Such compositions are prepared in accordance with acceptablepharmaceutical procedures, such as described in Remington'sPharmaceutical Sciences, 17th edition, ed. Alfonoso R. Gennaro, MackPublishing Company, Easton, Pa. (1985). Pharmaceutically acceptablecarriers are those that are compatible with the other ingredients in theformulation and biologically acceptable.

In another embodiment, the invention is directed to compositions,comprising:

a. a conjugate, comprising:

-   -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer;    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;        -   wherein said compound is covalently linked to said targeting            moiety; and    -   iii. a diagnostic or therapeutic metal; and        b. a pharmaceutically-acceptable carrier.

Pharmaceutically acceptable carriers, adjuvants, and vehicles that maybe used in the pharmaceutical compositions of this invention include,but are not limited to, ion exchangers, alumina, aluminum stearate,lecithin, serum proteins, such as human serum albumin, buffer substancessuch as phosphates, glycine, sorbic acid, potassium sorbate, TRIS(tris(hydroxymethyl)amino-methane), partial glyceride mixtures ofsaturated vegetable fatty acids, water, salts or electrolytes, such asprotamine sulfate, disodium hydrogen phosphate, potassium hydrogenphosphate, sodium chloride, zinc salts, colloidal silica, magnesiumtrisilicate, polyvinyl pyrrolidone, cellulose-based substances,polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,waxes, polyethylene-polyoxypropylene-block polymers, polyethyleneglycol, and wool fat.

According to this invention, the pharmaceutical compositions may be inthe form of a sterile injectable preparation, for example a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solutionand isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose, any bland fixed oil may be employed including synthetic mono-or di-glycerides. Fatty acids, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically-acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant.

In some cases, depending on the dose and rate of injection, the bindingsites on plasma proteins may become saturated with prodrug and activatedagent. This leads to a decreased fraction of protein-bound agent andcould compromise its half-life or tolerability as well as theeffectiveness of the agent. In these circumstances, it is desirable toinject the prodrug agent in conjunction with a sterile albumin or plasmareplacement solution. Alternatively, an apparatus/syringe can be usedthat contains the contrast agent and mixes it with blood drawn up intothe syringe; this is then re-injected into the patient.

The conjugates and pharmaceutical compositions of the present inventionmay be administered orally, parenterally, by inhalation spray,topically, rectally, nasally, buccally, vaginally or via an implantedreservoir in dosage formulations containing conventional non-toxicpharmaceutically-acceptable carriers, adjuvants and vehicles. The term“parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques.

When administered orally, the pharmaceutical compositions of thisinvention may be administered in any orally acceptable dosage formincluding, but not limited to, capsules, tablets, aqueous suspensions orsolutions. In the case of tablets for oral use, carriers that arecommonly used include lactose and corn starch. Lubricating agents, suchas magnesium stearate, are also typically added. For oral administrationin a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the activeingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening, flavoring or coloring agents may also beadded.

Alternatively, when administered in the form of suppositories for rectaladministration, the pharmaceutical compositions of this invention may beprepared by mixing the agent with a suitable non-irritating excipientthat is solid at room temperature but liquid at rectal temperature andtherefore will melt in the rectum to release the drug. Such materialsinclude cocoa butter, beeswax and polyethylene glycols.

As noted before, the pharmaceutical compositions of this invention mayalso be administered topically, especially when the target of treatmentincludes areas or organs readily accessible by topical application,including the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, poly-oxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith our without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

For administration by nasal aerosol or inhalation, the pharmaceuticalcompositions of this invention are prepared according to techniqueswell-known in the art of pharmaceutical formulation and may be preparedas solutions in saline, employing benzyl alcohol or other suitablepreservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other conventional solubilizing or dispersingagents.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. A typicalpreparation will contain from about 5% to about 95% active compound(w/w). Preferably, such preparations contain from about 20% to about 80%active compound.

For intravenous and other types of administration, acceptable doseranges range from about 0.001 to about 1.0 mmol/kg of body weight, withthe preferred dose of the active ingredient compound ranging from about0.001 to about 0.5 mmol/kg of body weight. Even more preferred is fromabout 0.01 to about 0.1 mmol/kg, and the most preferred dose of theactive ingredient compound is from about 0.02 and to about 0.05 mmol/kg.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage regimens for anyparticular patient will depend upon a variety of factors, including theactivity of the specific compound employed, the age, body weight,general health status, sex, diet, and time of administration, rate ofexcretion, drug combination and the judgment of the treating physician.

Kits

In yet other embodiments, the invention is directed to kits fordetecting, imaging, monitoring, or treating a disease or condition in apatient comprising:

-   a. a complex comprising:    -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   wherein said compound is covalently linked to said targeting        moiety;-   b. a therapeutic or diagnostic metal;-   c. an optional pharmaceutically-acceptable carrier; and-   d. instructions for preparing a composition comprising a therapeutic    or diagnostic agent for detecting, imaging, monitoring, or treating    a disease or condition in a patient.

The inclusion of one or more optional components in the kit willfrequently improve the ease of synthesis of the therapeutic anddiagnostic agent by practicing end user, the ease of manufacturing thekit, the shelf-life of the kit, or the stability and shelf-life of thediagnostic metal. The improvement achieved by the inclusion of anoptional component in the formulation must be weighed against the addedcomplexity of the kit and added cost to manufacture the kit. One or morevials that contain all or part of the formulation can independently bein the form of a sterile solution or a lyophilized solid.

Buffers useful in the preparation of the therapeutic and diagnosticagents and kits thereof include but are not limited to phosphate,citrate, sulfosalicylate, and acetate. A more complete list can be foundin the United States Pharmacopeia.

Lyophilization aids useful in the preparation of the therapeutic anddiagnostic agents and kits thereof include but are not limited tomannitol, lactose, sorbitol, dextran, Ficoll, and polyvinylpyrrolidine(PVP).

Stabilization aids useful in the preparation of therapeutic anddiagnostic agents and kits thereof include but are not limited toascorbic acid, cysteine, monothioglycerol, sodium bisulfite, sodiummetabisulfite, gentisic acid, and inositol.

Solubilization aids useful in the preparation of therapeutic anddiagnostic agents and kits thereof include but are not limited toethanol, glycerin, polyethylene glycol, propylene glycol,polyoxyethylene sorbitan monooleate, sorbitan monoloeate, polysorbates,poly(oxyethylene)-poly(oxypropylene)poly(oxyethylene) block copolymers(Pluronics) and lecithin. Preferred solubilizing aids are polyethyleneglycol, and Pluronics copolymers.

Bacteriostats useful in the preparation of therapeutic and diagnosticagents and kits thereof include but are not limited to benzyl alcohol,benzalkonium chloride, chlorbutanol, and methyl, propyl or butylparaben.

A component in a diagnostic kit can also serve more than one function. Areducing agent can also serve as a stabilization aid, a buffer can alsoserve as a transfer ligand, a lyophilization aid can also serve as atransfer, ancillary or coligand and so forth.

The predetermined amounts of each component in the formulation aredetermined by a variety of considerations that are in some casesspecific for that component and in other cases dependent on the amountof another component or the presence and amount of an optionalcomponent. In general, the minimal amount of each component is used thatwill give the desired effect of the formulation. The desired effect ofthe formulation is that the practicing end user can synthesize thetherapeutic or diagnostic agent and have a high degree of certainty thatthe therapeutic or diagnostic agent can be injected safely into apatient and will provide a therapeutic effect or diagnostic informationabout the disease state of that patient.

The kits of the present invention can also contain written instructionsfor the practicing end user to follow to synthesize the therapeutic ordiagnostic agents. These instructions may be affixed to one or more ofthe vials or to the container in which the vial or vials are packagedfor shipping or may be a separate insert, termed the package insert.

X-ray contrast agents and metallopharmaceuticals for magnetic resonanceimaging contrast agents are provided to the end user in their final formin a formulation contained typically in one vial, as either alyophilized solid or an aqueous solution. The end user reconstitutes thelyophilized solid with water or saline and withdraws the patient dose orsimply withdraws the dose from the aqueous solution formulation asprovided.

Methods of Use

In yet other embodiments, the invention is directed to kits fordetecting, imaging, monitoring, or treating a disease or condition in apatient comprising:

-   a. a conjugate comprising:    -   i. at least one targeting moiety selected from the group        consisting of peptide, protein, and amine-surface functional        dendrimer; and    -   ii. a residue of a compound of Formula I or Formula II or a        pharmaceutically acceptable salt thereof;    -   iii. a diagnostic or therapeutic metal;        -   wherein said compound is covalently linked to said targeting            moiety;-   b. an optional pharmaceutically-acceptable carrier; and-   c. instructions for preparing a composition comprising a diagnostic    agent for detecting, imaging, monitoring, or treating a disease or    condition in a patient.

In other embodiments, the invention is directed to methods of detecting,imaging or monitoring cancer in a patient, comprising the steps of:

-   a. administering to said patient a conjugate described above; and-   b. acquiring an image of a site of concentration of said conjugate    in the patient by a diagnostic imaging technique.    Preferable, the diagnostic imaging technique is magnetic resonance    imaging, single photon emission imaging, or positron emission    tomographic imaging.

In other embodiments, the invention is directed to methods of detecting,imaging or monitoring thrombi in a patient, comprising the steps of:

-   a. administering to said patient a conjugate described above; and-   b. acquiring an image of a site of concentration of said conjugate    in the patient by a diagnostic imaging technique.    Preferable, the diagnostic imaging technique is magnetic resonance    imaging, single photon emission imaging, or positron emission    tomographic imaging.

In yet other embodiments, the invention is directed to methods oftreating cancer, comprising the step of:

administering to a patient in need thereof an effective amount of aconjugate described above.

The present invention is further defined in the following Examples, inwhich all parts and percentages are by weight and degrees are Celsius,unless otherwise stated. It should be understood that these examples,while indicating preferred embodiments of the invention, are given byway of illustration only. From the above discussion and these examples,one skilled in the art can ascertain the essential characteristics ofthis invention, and without departing from the spirit and scope thereof,can make various changes and modifications of the invention to adapt itto various usages and conditions.

EXAMPLES Example 1 Preparation ofN—[(R)-2-amino-3-(p-nitrophenyl)propyl]trans-(S,S)-cyclohexane-1,2-diamine

N—[(R)-2-Amino-3-(p-nitrophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diaminewas previously generally prepared in accordance with Wu, C., Kobayashi,H., Sun, B., Yoo, T. M., Paik, C. H., Gansow, O. A., Carrasquillo, J.A., Pastan, I., Brechbiel, M. W.: Stereochemical Influence on theStability of Radio-Metal Complexes In Vivo. Synthesis and Evaluation ofthe Four Stereoisomers of 2-(p-nitrobenzyl)-trans-CyDTPA. Bioorg. Med.Chem. 1997, 5, 1925-1934. However, the procedure was modified in thatthe intermediate monoprotected 2,2-diaminocyclohexane was preparedaccording to a higher yielding route taken from Young. K. Kim, Seok J.Lee and Kyo H. Ahn: New hybrid ligands with atrans-1,2-diaminocyclohexane backbone: competing chelation modes inpalladium-catalysed enantioselective allylic alkylation. J. Org. Chem.2000, 65, 7807-7813.

Example 2 Preparation ofN—[(R)-2-amino-3-(p-nitrophenyl)propyl]trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N′,N″,N″-pentaethyl-t-butanoate

N—[(R)-2-Amino-3-(p-nitrophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine.3HCl (2 g, 5 mmol) stirred in acetonitrile (50 cm³) with K₂CO₃ (6.21 g,45 mmol) was treated with t-butylbromoacetate (5.17 cm³, 6.83 g, 35mmol) and stirred vigorously for 3 days. The solvent was evaporated atreduced pressure, ethylether (100 cm³) added and the mixture filtered.The inorganic salts were washed with additional portions of diethylether (3×10 cm³) and the filtrate evaporated at reduced pressure to aviscous orange oil. Purification was achieved on a silica gel column(previously treated with a mixture of water (10%) and ethanol (90%) thenrinsed with 100% ethanol followed by ethyl acetate) eluting with 30%ammonia in ethanol:ethyl acetate:hexane 1:1:16 to 1:1:8 gradient.

Example 3 Preparation ofN—[(R)-2-Amino-3-(p-aminophenyl)propyl]trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N′,N″,N″-pentaethyl-t-butanoate

10% palladium on carbon was placed in an all glass hydrogenation vesselwith ethanol (20 cm³) filled with hydrogen. After saturation of thecatalyst with hydrogen a solution ofN—[(R)-2-Amino-3-(p-nitrophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N′,N″,N″-pentaethyl-t-butanoate(2.7 g, 3.2 mmol) in ethanol (25 cm³) was injected and pressuremaintained at room pressure over 8 hours by periodic refilling ofhydrogen. The mixture was left vigorously stirring overnight thenfiltered through celite, washing with ethanol (5×5 cm³). The filtratewas evaporated at reduced pressure to give a pale yellow oil. Theproduct was purified on neutral alumina eluting with hexane:ethylacetate:NH₃ in ethanol 18:2:1 yielding a colorless oil afterevaporation.

Example 4 Preparation ofN—[(R)-2-Amino-3-(p-aminophenyl-N-{5-oxopentanoicacid}propyl]-trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N′,N″,N″-pentaethyl-t-butanoate

N—[(R)-2-Amino-3-(p-aminophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N′,N″,N″-pentaethyl-t-butanoate(1.85 g, 2.22 mmol), an excess of glutaric anhydride (0.507 g, 4.45mmol) were mixed in benzene (30 cm³) with stirring overnight. A furtherportion of benzene was added (30 cm³) and was washed with 0.1M Na₂HPO₃(1×30 cm³) followed by washing with 0.1M NaH₂PO₃. The organic fractionwas dried over Na₂SO₄, filtered then evaporated at reduced pressure toyield a pale brown oil. The product was further purified bychromatography on a silica gel column (previously treated with a mixtureof water (10%) and ethanol (90%) then rinsed with 100% ethanol followedby ethyl acetate) eluting with ethanol:hexane 1:5 to 1:1 gradient,yielding after evaporation at reduced pressure a glassy, colorlesssolid.

Example 5 Preparation ofN—[(R)-2-Amino-3-(p-5-[(2,5-dioxopyrrolidin-1-yl)oxy]-5-oxo-N-phenylpentanamide)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N″,N″-penta-t-butylacetateCompound of Formula I

1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.286 g,1.5 mmol), N-hydroxy succinimide (0.151 g, 1.3 mmol) andN—[(R)-2-Amino-3-(p-aminophenyl-N-{5-oxopentanoicacid})propyl]-trans-(S,S)-cyclohexane-1,2-diamine-N,N,N′,N″,N″-penta-t-butylacetate(1.13 g, 1.2 mmol) were stirred in a mixture of ethyl acetate (30 cm³)and DMF (10 cm³) overnight. The reaction mixture was then diluted withethyl acetate (30 cm³) and cooled in an ice bath. The cooled mixture waswashed with 5% w/v aqueous NaHCO₃ solution (2×20 cm³) ice cold and thenwith ice cold water (2×20 cm³). The organic fraction was dried overNa₂SO₄, filtered and evaporated at reduced pressure to yield a glassypale yellow solid. No further purification was performed on thiscompound.

Example 6 Preparation of Compound of Formula II

A suspension of an α-Fmoc-lysine (0.024 g) in dimethyl formamide (1 cm³)was treated with the compound of Formula I from Example 5 over a periodof 30 minutes the solution cleared. The mixture was stirred overnightthen ethyl acetate (20 cm³) added and the mixture washed with 0.1 MNaH₂PO₃ (4×10 cm³).

Example 7 Complex Formation

The compound of Formula I was tested as a bioconjugation reagent with asample of octatreotide. A mass spectrum of the complex formed indicatedsuccessful labeling of the unprotected amine terminus to form thecomplex of the compound of Formula I and octatreotide. NMR spectra ofthe unlabeled and labeled complex also confirmed complex formation.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges specific embodiments thereinare intended to be included.

The disclosures of each patent, patent application and publication citedor described in this document are hereby incorporated herein byreference, in its entirety.

Those skilled in the art will appreciate that numerous changes andmodifications can be made to the preferred embodiments of the inventionand that such changes and modifications can be made without departingfrom the spirit of the invention. It is, therefore, intended that theappended claims cover all such equivalent variations as fall within thetrue spirit and scope of the invention.

1. A compound of Formula I or II:

or a pharmaceutically acceptable salt thereof.
 2. A complex, comprising:a. at least one targeting moiety selected from the group consisting ofpeptide, protein, and an amine-surface functional dendrimer; and b. aresidue of a compound according to claim 1 or a pharmaceuticallyacceptable salt thereof; wherein said compound is covalently linked tosaid targeting moiety.
 3. A complex according to claim 2, wherein saidtargeting moiety is PAMAM dendrimer, octatreotide, bombesin, orα-melanocyte stimulating hormone.
 4. A complex according to claim 2,wherein said compound is linked to said targeting moiety via a terminalamino group on said targeting moiety.
 5. A complex according to claim 4,wherein said compound is linked to said targeting moiety via anN-terminus, a lysine residue, or shell amine.
 6. A complex according toclaim 2, wherein said compound is a residue of a compound of Formula Ior a pharmaceutically acceptable salt thereof.
 7. A complex according toclaim 2, wherein said compound is a residue of a compound of Formula IIor a pharmaceutically acceptable salt thereof.
 8. A conjugate,comprising: a. a complex according to claim 2 or a pharmaceuticallyacceptable salt thereof; and b. a diagnostic or therapeutic metal.
 9. Aconjugate according to claim 8, wherein said diagnostic or therapeuticmetal is selected from the group consisting of a paramagnetic metal ion,a gamma-emitting radioisotope, a positron-emitting radioisotope, and anx-ray absorber.
 10. A conjugate according to claim 9, wherein saidparamagnetic metal ion is selected from the group consisting of:Gd(III), Dy(III), Fe(III), and Mn(II).
 11. A conjugate according toclaim 9, wherein said diagnostic or therapeutic metal is agamma-emitting radioisotope or positron-emitting radioisotope selectedfrom the group consisting of: ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, ^(99m)Tc,⁹⁵Tc, ¹¹¹In, ⁹⁰Y, ¹⁴⁹Pr, ¹⁵³Sm, ¹⁵⁹Gd, ¹⁶⁶Ho, ¹⁶⁹Yb, ¹⁷⁷Lu, ¹⁸⁶Re,¹⁸⁸Re, and ²¹³Bi.
 12. A conjugate according to claim 11, wherein saiddiagnostic or therapeutic metal is ^(99m)Tc.
 13. A conjugate accordingto claim 11, wherein said diagnostic or therapeutic metal is ¹¹¹In. 14.A conjugate according to claim 11, wherein said diagnostic ortherapeutic metal is ²¹³Bi.
 15. A conjugate according to claim 9,wherein said x-ray absorber is a metal selected from the groupconsisting of: Re, Sm, Ho, Lu, Pm, Y, Bi, Pd, Gd, La, Au, Au, Yb, Dy,Cu, Rh, Ag, and Ir.
 16. A composition, comprising: a. a compoundaccording to claim 1 or a pharmaceutically acceptable salt thereof; andb. a pharmaceutically-acceptable carrier.
 17. A composition, comprising:a. a complex according to claim 2 or a pharmaceutically acceptable saltthereof; and b. a pharmaceutically-acceptable carrier.
 18. Acomposition, comprising: a. a conjugate according to claim 8 or apharmaceutically acceptable salt thereof; and b. apharmaceutically-acceptable carrier.
 19. A kit for detecting, imaging,monitoring, or treating a disease or condition in a patient comprising:a. a complex according to claim 2 or a pharmaceutically acceptable saltthereof; b. a therapeutic or diagnostic metal; c. an optionalpharmaceutically-acceptable carrier; and d. instructions for preparing acomposition comprising a therapeutic or diagnostic agent for detecting,imaging, monitoring, or treating a disease or condition in a patient.20. A kit for detecting, imaging, monitoring, or treating a disease orcondition in a patient comprising: a. a conjugate according to claim 8or a pharmaceutically acceptable salt thereof; b. an optionalpharmaceutically-acceptable carrier; and c. instructions for preparing acomposition comprising a therapeutic or diagnostic agent for detecting,imaging, monitoring, or treating a disease or condition in a patient.21. A process of synthesizing a peptide or protein comprising achelator, said process comprising the step of: incorporating into saidpeptide or protein a compound according to claim 1 or a pharmaceuticallyacceptable salt thereof.
 22. A process of synthesizing a peptide orprotein comprising a chelator, wherein said peptide or protein issynthesized by solid phase peptide synthesis.
 23. A peptide or proteinproduced by the process of claim 21 or claim
 22. 24. A process ofsynthesizing an amine-surface functional dendrimer comprising achelator, said process comprising the step of: incorporating into saidamine-surface functional dendrimer a residue of a compound according toclaim 1 or a pharmaceutically acceptable salt thereof.
 25. Anamine-surface functional dendrimer produced by the process of claim 24.26. A method of detecting, imaging or monitoring cancer in a patient,comprising the steps of: a. administering to said patient a conjugateaccording to claim 8 or a pharmaceutically acceptable salt thereof; andb. acquiring an image of a site of concentration of said conjugate inthe patient by a diagnostic imaging technique.
 27. A method according toclaim 26, wherein said diagnostic imaging technique is magneticresonance imaging, single photon emission imaging, or positron emissiontomographic imaging.
 28. A method of detecting, imaging or monitoringthrombi in a patient, comprising the steps of: a. administering to saidpatient a conjugate according to claim 8 or a pharmaceuticallyacceptable salt thereof; and b. acquiring an image of a site ofconcentration of said conjugate in the patient by a diagnostic imagingtechnique.
 29. A method according to claim 28, wherein said diagnosticimaging technique is magnetic resonance imaging, single photon emissionimaging, or positron emission tomographic imaging.
 30. A method oftreating cancer, comprising the step of: administering to a patient inneed thereof an effective amount of a conjugate according to claim 8 ora pharmaceutically acceptable salt thereof.
 31. A method of treatingthrombosis, comprising the step of: administering to a patient in needthereof an effective amount of a conjugate according to claim 8 or apharmaceutically acceptable salt thereof.